pPeOp from Omphalia lapidescens Schroet induces cell cycle arrest and inhibits the migration of MC-4 gastric tumor cells.
pPeOp from Omphalia lapidescens Schroet induces cell cycle arrest and inhibits the migration of MC-4 gastric tumor cells.
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来自 Omphalia lapidescens Schroet 的 pPeOp 诱导细胞周期停滞并抑制 MC-4 胃肿瘤细胞的迁移
DOI：
10.3892/ol.2017.6207
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发表时间：
2017-07
期刊：
影响因子：
2.9
通讯作者：
Chen YT
中科院分区：
文献类型：
--
作者：
Yang YL;Gong WY;Chen FF;Chen LC;Chen YT
本研究旨在研究用聚乙烯吡啶烷酮从石蜡菌中提取纯化石蜡蛋白（pPeOp）对MC-4人胃癌细胞增殖和细胞周期进程的影响。以聚乙烯吡咯烷酮为原料，从石蜡中提取pepep。MC-4细胞分别用30、60或90µg/ml pep培养，5-氟尿嘧啶作为阳性对照。与未处理的对照组相比，处理后的细胞存活率显著降低，且呈剂量依赖性。流式细胞术分析显示，经pep处理的细胞停留在S期，并表现出异常的G0/G1和G2/M期细胞周期分布。此外，伤口愈合实验表明，pep显著抑制MC-4细胞的迁移。采用逆转录-定量聚合酶链式反应和western blotting检测细胞周期蛋白D1/细胞周期蛋白依赖性激酶（CDK） 4、细胞周期蛋白B/CDK1、细胞周期蛋白A/CDK2、基质金属蛋白酶(MMP)-2和MMP-9 mRNA和蛋白表达水平。CDK4、细胞周期蛋白A mRNA表达水平较未处理对照组显著升高；而cyclin D1、CDK1、CDK2、cyclin B、MMP-2、MMP-9 mRNA表达水平显著降低，表明浓度与cyclin D1表达水平呈负相关。周期阻滞相关蛋白和迁移相关蛋白的表达与观察到的mRNA表达水平相似。综上所述，在MC-4细胞中，pPeOp可以抑制迁移并导致S期细胞周期阻滞。
The aim of the present study was to investigate the effect of purified Omphalia lapidescens protein (pPeOp) extracted by polyvinylpyrrolidone from the fungus Omphalia lapidescens Schroet on the proliferation and cell cycle progression of MC-4 human gastric tumor cells. Using polyvinylpyrrolidone, pPeOp was extracted from O. lapidescens Schroet. MC-4 cells were cultured with 30, 60 or 90 µg/ml pPeOp, with 5-fluorouracil used as a positive control. Survival rates of treated cells were significantly decreased compared with those of the untreated control group in a dose-dependent manner. Using flow cytometric analysis, cells treated with pPeOp were demonstrated to arrest in S phase and exhibit abnormal G0/G1 and G2/M phase cell cycle distribution. In addition, a wound healing assay demonstrated that pPeOp significantly inhibited the migration of MC-4 cells. The mRNA and protein expression levels of cyclin D1/cyclin-dependent kinase (CDK) 4, cyclin B/CDK1, cyclin A/CDK2, matrix metalloproteinase (MMP)-2 and MMP-9 were determined using reverse transcription-quantitative polymerase chain reaction analysis and western blotting. The mRNA expression level of CDK4 and cyclin A was significantly increased compared with the untreated control; however, cyclin D1, CDK1, CDK2, cyclin B, MMP-2, and MMP-9 exhibited a significantly decreased mRNA expression level, indicating that there is a negative association between concentration and cyclin D1 expression levels. The expression of the cycle arrest-associated proteins and migration-associated proteins examined were similar to the observed mRNA expression levels. In conclusion, pPeOp was identified to inhibit migration of and cause S phase cell cycle arrest in MC-4 cells.